title: Sperm-associated antigen 11A is expressed exclusively in the principal cells of the mouse caput epididymis in an androgen-dependent manner.
abstract: BACKGROUND
  Epididymal sperm maturation occurs via interactions between sperm and proteins secreted by the epididymal epithelium. Although this is an important process, the genes that encode the involved proteins remain largely uncharacterized. Previous studies have demonstrated that the genes involved in sperm maturation are regulated by androgen. Spag11a is an epididymal gene that is influenced by androgen. However, little is known about the putative role of this gene in the sperm maturation process. The objective of this study was to characterize Spag11a in the mouse epididymis.
  
  
  METHODS
  In silico analyses were performed to predict signal peptides and functional domains. Spag11a expression was measured by quantitative real-time RT-PCR. Western blots and immunocytochemistry were performed to determine protein expression.
  
  
  RESULTS
  SPAG11A is a member of the beta defensin protein family and constitutes a secretory protein. Spag11a was expressed exclusively in the epididymis. Moreover, it exhibited region-specific expression in the caput, which is typical for genes that are involved in creating a suitable microenvironment for sperm maturation. Mouse Spag11a was regulated by androgen. A significant decrease of Spag11a expression was observed at third day following a gonadectomy (P < 0.001). Interestingly, testosterone replacement therapy was able to maintain the expression almost at the normal level, indicating a dependency on androgen. Besides androgen, testicular factors influenced Spag11a expression in a different way. This was revealed by efferent duct ligation in which Spag11a was transiently up-regulated at the third day following the ligation before returning to the normal level at day 5. Spag11a regional expression was also observed at protein level detected by western immunoblotting which revealed a clear band in the caput but not in other regions. The prediction that SPAG11A is a secretory protein was confirmed by immunocytochemical analyses indicating cell-specific expression mainly in the caput principal cells and detection of the protein in epididymal luminal fluid and spermatozoa.
  
  
  CONCLUSIONS
  Based on the characteristics of Spag11a, it is likely that this gene has a specific role in epididymal sperm maturation. Further studies using functional assays are necessary to confirm this finding.
authors.affiliation: Department of Biology, Faculty of Medicine, University of Indonesia, Jl, Salemba Raya 6, Jakarta 10430, Indonesia. dwi.ari@ui.ac.id
authors.forename: Dwi A
authors.initials: DA
authors.lastname: Pujianto
authors.forename: Evelyn
authors.initials: E
authors.lastname: Loanda
authors.forename: Puji
authors.initials: P
authors.lastname: Sari
authors.forename: Yurnadi H
authors.initials: YH
authors.lastname: Midoen
authors.forename: Purnomo
authors.initials: P
authors.lastname: Soeharso
mesh_terms: D000595:Amino Acid Sequence; D000728:Androgens / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D004822:Epididymis / Q000166:cytology / Q000378:metabolism*; D020869:Gene Expression Profiling; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D009919:Orchiectomy; D010766:Phosphorylation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013079:Sperm Maturation / Q000235:genetics; D013094:Spermatozoa / Q000378:metabolism; D013997:Time Factors; D023083:beta-Defensins / Q000235:genetics* / Q000378:metabolism
chemical_list: D000728:Androgens; C465595:Spag11 protein, mouse; D023083:beta-Defensins